RNA forms various secondary structures depending on its sequence in cells; the influence of secondary structures must be considered when detecting, regulating, or manipulating RNA. In this study, we utilized the cooperativity of multiple photo‐cross‐linkable assistant probes to evaluate photo‐cross‐linking for RNAs with complex secondary structures. We succeeded in increasing the fluorescence intensity 39.9 times in the region where detection was difficult owing to the formation of a complex secondary structure in the conventional fluorescence in situ hybridization method. This method enables the detection, regulation, and manipulation of RNA that forms various structures depending on the sequence.
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